Immunological Electrospun Scaffold For Tumor Imprison Killing And Healthy Tissue Regeneration

Objective: We fabricate the multifunctional scaffolds with suppressing tumor,activating immune response and promoting tissue regeneration properties.Methods: The poly(L-lactide)(PLLA)electrospun fibrous scaffolds were prepared by electrospinning.Agonist mouse anti-human CD40 antibody(CD40mAb)is grafted to the surface of the PLLA electrospun fibrous scaffolds through dopamine(PDA)mediated crosslinking reaction under mild aqueous condition.(1)The physical and chemical properties of PLLA,PLLA-PDA,PLLA-PDA-IgG and PLLA-PDA-CD40 mAb were explored by X-ray photoelectron spectroscopy(XPS)and scanning electron microscopy(SEM),respectively.The tensile stress of the electrospun fibrous scaffolds were tested by mechanical tensile test.The hydrophilic properties of the electrospun fibrous scaffolds by water contact angle(WCA).(2)The absorbed and released profile of CD40 mAb on PLLA-PDA scaffolds.(3)Cell viability of MDA-MB-231 is evaluated with CCK-8 kit.The relative gene expressions of Bax and Bcl-2 of MDA-MB-231 cells are quantified by qRT-PCR.Flow cytometry assay for MDA-MB-231 cell apoptosis.(4)Flow cytometry assay was used to detect the ability of induced maturation of DCs.The morphology of DC cells was observed by microscope.(5)Cell viability of MC3T3-E1 is evaluated with CCK-8 kit.Results:(1)XPS data showed successful grafting of PDA and antibodies,the diameters of PLLA,PLLA-PDA,PLLA-PDA-IgG,PLLA-PDA-CD40 mAb fiber were similar.,WCA proved that the scaffolds changed from hydrophobic into hydrophilic surface with PLLA grafting PDA motif,which benefit to cell attachment.Mechanical tensile test showed that the mechanical properties of fiber scaffolds were not changed before and after grafting.(2)The fluorescent signal is observed in PLLA-PDA electrospun fibrous scaffolds conjugated with FITC-labeled antibodies,indicating the successful grafting of IgG or CD40 mAb onto the scaffolds.The absorption curve showed that CD40 mAb could be completely grafted to the fibrous scaffold for 24 h.The release curve indicated that PLLA-PDA-CD40 mAb could release CD40 mAb with 48 h.(3)CCK-8 proved that CD40 mAb released from PLLA-PDA-CD40 mAb could effectively inhibit the growth of human breast cancer cell MDA-MB-231.FCA and qRT-PCR showed that PLLA-PDA-CD40 mAb could induce cells apoptosis.With the fluorescent microscope,the images display more red cells(dead cells)on PLLA-PDA-CD40 mAb scaffolds.(4)We display that PLLA-PDA-CD40 mAb scaffolds have capability to promote the maturation of DCs by offering CD40 signaling.(5)All the results indicate that the CD40 mAb released from the scaffolds do not influence the growth of MC3T3-E1 cells.Conclusion: The developed PLLA-PDA-CD40 mAb scaffold is multifunctional in antitumor immunity and healthy tissue regeneration,which provides an alternative and promising strategy in cancer treatment.